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- EMDB-64623: Cryo-EM structure of the 48-nm repeat doublet microtubule from Te... -

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Basic information

Entry
Database: EMDB / ID: EMD-64623
TitleCryo-EM structure of the 48-nm repeat doublet microtubule from Tekt1 KO mouse sperm
Map data
Sample
  • Complex: sperm flagellar doublet microtubules
Keywordssperm flagellar doublet microtubules Tekt1 / STRUCTURAL PROTEIN
Biological speciesMus musculus (house mouse)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.8 Å
AuthorsLiu Q / Gui M / Zhou LN
Funding support China, 1 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC) China
CitationJournal: Nat Commun / Year: 2026
Title: Doublet microtubule-associated tektins and enzymes differentially regulate sperm flagellar integrity and motility.
Authors: Qi Liu / Lunni Zhou / Xiaochen Liang / Pengyu Chen / Bo Li / Shuo Yang / Yuqi Liu / Haibo Zhao / Jin Hu / Shan Feng / Shanshan Xie / Jianping Wu / Miao Gui /
Abstract: Doublet microtubule (DMT)-associated proteins assemble and drive sperm flagella, which are essential for successful fertilization. However, the exact roles of different DMT-associated proteins in ...Doublet microtubule (DMT)-associated proteins assemble and drive sperm flagella, which are essential for successful fertilization. However, the exact roles of different DMT-associated proteins in sperm function and the underlying molecular mechanisms remain elusive. Here, we generate four gene-knockout mice based on high-resolution structures targeting distinct DMT components: two intermediate filament-like tektins (TEKT1, TEKT5) and two enzymes (TSSK6, DUSP21). The depletion of TEKT1, shared by sperm flagella and motile cilia, causes male infertility characterized by impaired sperm motility and loss of the tektin bundle, whereas sperm-specific Tekt5 knockout (KO) mice remain fertile with largely normal flagellar function, indicating functional divergence within the tektin family. Tssk6 KO spermatozoa exhibit severely disturbed morphology and motility, resulting in homozygote infertility and heterozygote subfertility. Phosphoproteomics reveals dysregulated phosphorylation of axonemal proteins, highlighting the critical role of kinase-mediated signaling in regulating sperm motility. Conversely, Dusp21 KO mice display no fertility or sperm motility defects, suggesting compensatory phosphatase activity. Phenotypic comparisons between Tekt1 and Tssk6 KO mice suggest their involvement in distinct subtypes of asthenozoospermia. Overall, this study elucidates how filamentous and enzymatic DMT proteins govern sperm function through divergent mechanisms, which have implications for molecular diagnosis of male infertility.
History
DepositionMay 15, 2025-
Header (metadata) releaseMar 18, 2026-
Map releaseMar 18, 2026-
UpdateMar 18, 2026-
Current statusMar 18, 2026Processing site: PDBc / Status: Released

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Structure visualization

Supplemental images

emd_64623.png

Downloads & links

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Map

FileDownload / File: emd_64623.map.gz / Format: CCP4 / Size: 1.7 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.93 Å/pix.
x 764 pix.
= 710.52 Å		0.93 Å/pix.
x 764 pix.
= 710.52 Å		0.93 Å/pix.
x 764 pix.
= 710.52 Å

Surface

Projections

Slices (1/3)

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.93 Å
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Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.004
Minimum - Maximum-0.00823132 - 0.0151986135
Average (Standard dev.)0.00009244179 (±0.0009906781)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions764764764
Spacing764764764
CellA=B=C: 710.52 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: raw map

Fileemd_64623_additional_1.map
Annotationraw map
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AxesZYX

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Half map: #1

Fileemd_64623_half_map_1.map
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Half map: #2

Fileemd_64623_half_map_2.map
Projections & Slices
AxesZYX

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Sample components

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Entire : sperm flagellar doublet microtubules

EntireName: sperm flagellar doublet microtubules
Components
  • Complex: sperm flagellar doublet microtubules

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Supramolecule #1: sperm flagellar doublet microtubules

SupramoleculeName: sperm flagellar doublet microtubules / type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Mus musculus (house mouse)

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation statefilament

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Sample preparation

BufferpH: 7.2
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 1.25 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: EMDB MAP
EMDB ID:

35823

Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 26075
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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